Peptides having pharmacological activity for treating disorders associated with altered cell migration, such as cancer

ABSTRACT

Peptides and their functionally equivalent derivatives, in salified or non-salified form, with the general formula L 1 -X 1 —X 2 —X 3 —X 4 , wherein: 
     L 1  is H, or acyl, or any natural or non-natural amino acid, optionally N-acylated, N-alkylated and/or Cα-alkylated;
 
X 1  and X 3 , which are equal or different, are any natural or non-natural basic amino acid, optionally N-alkylated and/or Cα-alkylated;
 
X 2  is any natural or non-natural amino acid, optionally N-alkylated and/or Cα-alkylated, with the proviso that it is not glycine and amino acids mono-substituted on the α carbon atom with a linear or cyclic alkyl group, from 1 to 10 carbon atoms, and amino acids mono-substituted on the α carbon atom with a linear or cyclic alkyl group containing 4 to 10 carbon atoms, or amino acids mono-substituted on the α carbon atom with an alkyl group containing 1 to 8 carbon atoms, optionally substituted with a carbamoyl, hydroxyl or aromatic group;
 
X 4  is any natural or non-natural hydrophobic amino acid, optionally Cα-alkylated and/or amidated at the C-terminal end, or any hydrophobic amino alcohol, or a hydrophobic gem-diamine, optionally N′-alkylated or N′-acylated.

This invention relates to linear peptides and their functionallyequivalent derivatives, whether salified or unsalified, containing fouror five amino acid residues, at least one of which is hydrophobic and atleast two of which are basic, hereinafter called “PICM” (PeptideInhibitors of Cell Motility) and the pharmaceutical compositionscontaining them as active ingredients. The compounds according to theinvention are effective in the treatment and prevention of tumours,especially those which are highly invasive and/or liable to metastatise,and in the treatment of disorders connected with neo-angiogenesis andneo-vascularisation, and those associated with altered cell motilitysuch as autoimmune diseases and chronic inflammatory disorders likerheumatoid arthritis and psoriasis, chronic granulomatous disorders,retinopathies, macular degeneration and oedema, Kaposi's sarcoma anddiseases associated with herpes virus infection.

BACKGROUND OF THE INVENTION

Current tumour treatments are limited by the existence of highlymalignant cell types, which intrinsically fail to respond toconventional treatments (gliomas, sarcomas, etc.), or by the onset ofselective advantages, which promote the selection and consequentproliferation of resistant tumour clones during treatment (Woodhouse, E.C., et al., Cancer 80:1529-1537, 1997). Conventional treatments aremainly designed to inhibit the growth of the tumour rather thanpreventing its metastatic spread, which is still the main cause oftreatment failures (Shevde L A et al. Cancer Lett, 198:1-20, 2003).

The general characteristic of current tumour treatments is the use ofhighly cytotoxic compounds which, although they act selectively onmalignant cells, inevitably have devastating systemic effects on thebody. The result is that current treatments generally involve very highsocial, human and financial costs.

This overall picture demonstrates that: a) there is a pressing need todevelop effective treatments for currently untreatable tumours; b) thereis a strong need to reduce the side effects which make patients' qualityof life unacceptable when they are treated with current anti-tumourdrugs, and can even cause death in debilitated patients; c) the efficacyof treatments needs to be improved by using drugs that interfere bothwith the growth process and with the metastatic spread of the tumour; d)the cost of tumour treatments needs to be made more acceptable.

It was recently reported that the numerous biological actions of thepeptide Metastin, of human and murine derivation (WO 00/24890, WO01/75104, WO 02/85399) include an effect in the prevention or treatmentof cancer. WO 06/001499, which relates to Metastin and its derivatives,claims a very broad series of compounds, estimated at over 10¹⁰different structures, containing 4 to 54 amino acid residues, naturaland non-natural, for which a very wide variety of biological activity isreported, such as inhibition of metastatic spread and growth of tumours,control of the pancreatic function and prevention of acute and chronicpancreatitis, control of the placental function and use in the treatmentof foetal hypoplasia, abnormal glucose metabolism, abnormalities in thelipid metabolism, infertility, endometriosis, premature puberty,Alzheimer's disease, disorders affecting the cognitive sphere, obesity,hyperlipidaemia, diabetes mellitus type II, hyperglycaemia,hypertension, diabetic neuropathies, diabetic nephropathies, diabeticretinopathies, oedema, urinary disorders, insulin resistance, unstablediabetes, fatty atrophy, insulin allergies, atherosclerosis, thromboticdisorders, lipotoxicity, and use in treatments to improve the functionof the gonads and stimulate ovulation. The simultaneous existence ofsuch different biological actions for each of these compounds certainlyrepresents a major limitation, not an advantage, with a view to thetherapeutic application of this class of molecules. This wide variety ofbiological functions is closely associated with the interaction ofMetastin and its derivatives with the specific cell receptor GPR54, alsoknown as Kiss-1R, Kisspeptins receptor, Metastin receptor,hypogonadotropin 1 or hOT7T175 (Ohtaki T., et al., Nature 411:613-617,2001; Clements M. K., et al., Biochem. Biophys. Res. Commun.284:1189-1193, 2001; Muir A. I., et al., J. Biol. Chem. 276:28969-28975;2001; Kotani M., et al., J. Biol. Chem. 276:34631-34636, 2001; SeminaraS. B., et al., N. Engl. J. Med. 349:1614-1627, 2003; Grimwood J., etal., Nature 428:529-535, 2004; Colledge W. H., Trends Endocrinol. Metab.15:448-453, 2004; Hori A., et al., Biochem. Biophys. Res. Commun.286:958-963, 2001; Janneau J.-L., et al., J. Clin. Endocrinol. Metab.87:5336-5339, 2002; Ringel M. D., et al., J. Clin. Endocrinol. Metab.87:2399-2399, 2002; de Roux N., et al., Proc. Natl. Acad. Sci. U.S.A.100:10972-10976, 2003; Ikeguchi M., et al., J. Cancer Res. Clin. Oncol.129:531-535, 2003; Ikeguchi M., et al., Clin. Cancer Res. 10:1379-1383,2004; Bilban M., et al., J. Cell Sci. 117:1319-1328, 2004; Becker J. A.J., et al., Biochem. Biophys. Res. Commun. 326:677-686, 2005; Semple R.K., et al., J. Clin. Endocrinol. Metab. 90:1849-1855, 2005).

As regards the anti-tumoral activity of Metastin and its derivatives, WO06/001499 reports a modest activity, limited to experimental animalmodels, at the dose of 70-140 μg/kg, which reduces the tumour mass bynot more than 20%. It has also been demonstrated that long-termadministration of Kisspeptin-54, one of the analogues of Metastin,causes the adverse effect of atrophy of the gonads in the rat (E. L.Thomson et al., Am. J. Physiol. Endocrinol. Metab. 291, 1074-1082,2006).

DESCRIPTION OF THE INVENTION

This invention concerns tetra-peptides and penta-peptides and theirfunctionally equivalent derivatives, in salified or unsalified form,containing at least two basic and at least one hydrophobic amino acids(hereinafter indicated by the code PICM), which inhibit cell migrationin vitro at aM concentrations (10⁻¹⁸M), and have a powerful antitumoralaction in vivo, reducing the tumour mass from 30 to 70% at the dose aslow as 15 μg/kg. They are effective in all disorders associated withneo-angiogenesis and neo-vascularisation, and do not present any acuteor sub-acute toxicity up to doses approx. 1000 times higher than thetherapeutic dose.

Unlike Metastin and its derivatives, the activity of the peptidesaccording to the invention is not correlated with cell receptor GPR54,but is strictly correlated with their specific interaction with the FPRcell receptors.

Some FPR receptor antagonists are known (Edwards et al. Mol Pharmacol.68:1301-10, 2005; Karisson et al. Biochem Pharmacol. 71:1488-96, 2006),all of which are chemically distinct from the PICMs.

The PICMs therefore represent a new class of active constituents whichpossess biological and pharmacological activities that are far morepotent than and different from those of Metastin and its derivatives andthe FPR receptor antagonists.

In clinical practice, PICMs are effective against a large number oftumours, act at low doses, and do not present any systemic toxicity oradverse effects.

The PICMs are also effective in the treatment of disorders connectedwith neo-angiogenesis and neo-vascularisation, and those associated withaltered cell motility such as autoimmune diseases and chronicinflammatory disorders like rheumatoid arthritis and psoriasis, chronicgranulomatous disorders, retinopathies, macular degeneration and oedema,Kaposi's sarcoma and diseases associated with herpes virus infection.

DETAILED DESCRIPTION OF THE INVENTION

The peptides according to the invention and their functionallyequivalent derivatives, in salified or non-salified form, have thegeneral formula L₁-X₁—X₂—X₃—X₄, wherein: L₁ is H, or acyl, or anynatural or non-natural amino acid, optionally N-acylated or N-alkylatedand/or Cα-alkylated; preferably, L₁ is H, or acyl, or Glu, Gln,optionally N-acylated or N-alkylated and/or Cα-alkylated, Pro,hydroxy-Pro, thio-Pro, Azt, Pip, pGlu, optionally N-acylated and/orCα-alkylated, Aib, Ac3c, Ac4c, Ac5c, Ac6c, optionally N-acylated orN-alkylated; even more preferably L₁ is H or acyl, Aib, Ac3c, Ac4c, Ac5cor Ac6c, optionally N-acylated or N-alkylated;

X₁ and X₃, which are equal or different, are any natural or non-naturalbasic amino acid, optionally N-alkylated and/or Cα-alkylated;preferably, X₁ and X₃, which are equal or different, optionallyN-alkylated and/or Cα-alkylated, are chosen from among Arg, Orn, Lys,optionally guadinylated, and phenylalanine substituted in the meta orpara positions with an amine or guanidine group;X₂ is any natural or non-natural amino acid, optionally N-alkylatedand/or Cα-alkylated, with the proviso that it is not glycine and aminoacids mono-substituted on the α carbon atom with a linear or cyclicalkyl group, from 1 to 10 carbon atoms, or of amino acids monosubstituted on the α carbon atom with a linear or cyclic alkyl groupcontaining 4 to 10 carbon atoms, or amino acids mono-substituted on theα carbon atom with an alkyl group containing 1 to 8 carbon atoms,optionally substituted with a carbamoyl, hydroxyl or aromatic group; X₂is preferably chosen from among Glu, Lys, optionally N-alkylated and/orCα-alkylated, Aib, Ac3c, Ac4c, Ac5c, and Ac6c, optionally N-alkylated;more preferably, X₂ is chosen from among Aib, Ac3c, Ac4c, Ac5c and Ac6c,optionally N-alkylated;X₄ is any hydrophobic amino acid, optionally Cα-alkylated and/oramidated at the C-terminal end, or any hydrophobic amino alcohol or anyhydrophobic gem-diamine, optionally N′-alkylated or N′-acylated; X₄ ispreferably chosen from among Phe, h-Phe, Tyr, Trp, 1-NaI, 2-NaI,h-1-NaI, h-2-NaI, Cha, Chg and Phg, optionally Cα-alkylated and/oramidated at the C-terminal end.

“Natural amino acids” refers to the amino acids constituting the proteinof living organisms.

“Non-natural amino acids” refers to: α-amino acids in the D series orβ-amino acids; dehydro-amino acids; amino acids di-substituted on the αcarbon atom with alkyl or aryl groups containing up to 11 carbon atoms;natural amino acids, as defined above, containing, on the side chain,hydroxy, amino or thio groups functionalised with alkyls, acyls, arylsor acylaryls containing 1 to 11 carbon atoms; natural amino acids, asdefined above, containing, on the side chain, carboxy groupsfunctionalised with primary or secondary amines or aliphatic or aromaticalcohols containing up to 11 carbon atoms; cyclic amino acids such asAzt, thio-Pro, Ac3c, Ac4c, Ac5c, Ac6c, and pGlu acid; homo-amino acids;amino acids substituted by cycloalkyl or aryl groups such asβ-1-naphthyl-alanine, β-2-naphthyl-alanine, homo-β-1-naphthyl-alanine,homo-β-2-naphthyl-alanine, cyclohexyl-alanine, cyclohexyl-glycine, andphenyl-glycine. Other non-natural amino acids are those reported in:“Diversity of synthetic peptides”, Konishi et al. First InternationalPeptide Symposium, Kyoto, Japan, 1997.

The term “any basic amino acid” refers to any natural or non-naturalamino acid as defined above, containing at least one imidazole, amino,guanidino, pyridinium or urea group in the side chain.

The term “any hydrophobic amino acid” refers to α-, β- and dehydro-aminoacids in the series: Leu, n-Leu, Ile, allo-Ile, Val, n-Val, Phe, h-Phe,Tyr, Trp, 1-NaI, 2-NaI, h-1-NaI, h-2-NaI, Cha, Chg and Phg; natural andnon-natural amino acids as defined above, containing on the side chainhydroxy, amino or thio groups functionalised with alkyls, acyls, arylsor acylaryls containing 1 to 11 carbon atoms; natural and non-naturalamino acids, as defined above, containing on the side chain carboxygroups functionalised with primary or secondary amines or aliphatic oraromatic alcohols containing up to 11 carbon atoms; phenylalanines mono-and di-substituted in the ortho, meta and para positions of the aromaticring with halogens or with alkyl, O-alkyl or S-alkyl groups; β-2- andβ-3-thienylalanine, β-2- and β-3-furanylalanine; derivatives of 2,3di-amino propionic acid and 2,4 di-amino butyric acid functionalisedwith alkyls, acyls, aryls or acylaryls containing up to 11 carbon atoms.

The term “any hydrophobic amino alcohol” refers to “any hydrophobicamino acid” as defined above, wherein the carboxyl function issubstituted with an OH group.

The term “any hydrophobic gem-diamine” refers to “any hydrophobic aminoacid” as defined above, wherein the carboxyl function is substitutedwith an NH₂ group.

The term “acyl” means an acyl group containing 1 to 9 carbon atoms.

The term “N-acylated” means the introduction of an acyl, as definedabove, onto the terminal amino nitrogen.

The term “N-alkylated” means the introduction of an alkyl residuecontaining 1 to 9 carbon atoms onto the amide nitrogen.

The term “amidated” means the amidation of the C-terminal carboxyl witha primary or secondary amine containing a total of 0 to 14 carbon atoms.

The term “Cα-alkylated” means the introduction onto the α carbon atom ofan alkyl residue containing 1 to 9 carbon atoms.

The term “functionally equivalent derivatives” means derivatives of thecompounds with general formula I characterised by structuralmodifications conventionally used in peptide chemistry to modulate theirpharmacodynamic or pharmacokinetic properties. These includepseudo-peptides wherein one or more peptide bonds are substituted by—CH₂—NH— (Guichard G, et al., J Biol Chem.; 270:26057-9 1995),derivatives with one or more inverted peptide bonds (Carotti A., et al.Biopolymers 60, 322-332, 2001; Chorev, M., et al. Science 204, 1210-12121979; Pallai, P., et al. Biochemistry 24, 1933-1941. 1985; Rodriguez M.E. et al. J. Med. Chem. 30, 758-763 1987), β-peptide derivatives (HorneW. S. et al. J. Am. Chem. Soc. 129 4178-4180 2007), wherein one or morepeptide bonds are formed by at least one β-amino acid, and derivativescontaining one or more dehydro-amino acids (Busetti V. et al. Int. J.Biol. Macromol. 14, 23-28 1992). Derivatives with elongation of thechain from the N-terminal side are also functionally equivalentderivatives.

The following are the conventional abbreviations used for some of thenon-natural amino acids which can be included in the formulas of thepeptides according to the invention:

Azt=azetidinic acid, Pip=pipecolic acid, Aib=α-amino-isobutyric acid,Ac3c=1-aminocyclopropan-1-carboxylic acid,Ac4c=1-aminocyclobutan-1-carboxylic acid,Ac5c=1-aminocyclopentan-1-carboxylic acid,Ac6c=1-aminocyclohexan-1-carboxylic acid, Abu=α-amino-n-butyric acid,n-Leu=norleucine, n-Val=norvaline, h-Phe=homo-phenylalanine,1-NaI=β-1-naphthyl-alanine, 2-NaI=β-2-naphthyl-alanine,h-1-NaI=homo-β-1-naphthyl-alanine, h-2-NaI=homo-β-2-naphthyl-alanine,Cha=cyclohexyl-alanine, Chg=cyclohexyl-glycine, Phg=phenyl-glycine,pGlu=pyroglutamic acid, Dap=2,3 di-amino-propionic acid, Dab=2,4diaminobutyric acid, N(Me)Arg=N-methyl-arginine,α(Me)Phe=C-alpha-methyl-phenylalanine.

Preferably, L1 is acetyl, Glu, pGlu, acetyl-Aib, X1 is Arg or N(Me)Arg,X2 is Glu, Aib, Ac5c, X3 is Arg, N(Me)Arg, and X4 is Phe-NH₂, Tyr-NH₂,Trp-NH₂, α(Me)Phe-NH₂, Phe-OH, Tyr-OH, Trp-OH.

The particularly preferred peptides according to the invention arechosen from among: Ace-Arg-Glu-Arg-Phe-NH₂;Ace-Arg-Glu-N(Me)Arg-Phe-NH₂; Ace-Arg-Aib-Arg-Phe-NH₂;Ace-Arg-Aib-N(Me)Arg-Phe-NH₂; Ace-Arg-Ac5c-Arg-Phe-NH₂;Ace-Arg-Ac5c-N(Me)Arg-Phe-NH₂; Ace-N(Me)Arg-Aib-Arg-Phe-NH₂;Ace-N(Me)Arg-Aib-N(Me)Arg-Phe-NH₂; Ace-Arg-Aib-N(Me)Arg-Phe-NH₂;Ace-Arg-Aib-Arg-α(Me)Phe-NH₂; Ace-N(Me)Arg-Aib-Arg-α(Me)Phe-NH₂;Ace-N(Me)Arg-Aib-N(Me)Arg-α(Me)Phe-NH₂;Ace-Arg-Aib-N(Me)Arg-α(Me)Phe-NH₂.

The peptides according to the invention can be synthesised with thevarious techniques reported in the literature; see, for example,Schroeder et al. “The Peptides” vol 1, Academic Press, 1965; Bodanszkyet al. “Peptide Synthesis Interscience Publisher, 1966; Barany &Merrifield, “The peptides; Analysis, Synthesis, Biology”, 2, Chapter 1,Academic Press, 1980. These techniques include solid-phase peptidesynthesis, peptide synthesis in solution, organic chemistry synthesismethods, or any combination thereof. The synthesis method chosen willobviously depend on the composition of the particular peptide.Preferably, the methods used will be based on appropriate combinationsof solid-phase techniques and classic methods in solution, which involvelow manufacturing costs, especially on an industrial scale. In detail,these methods involve: i) synthesis in solution of fragments of thepeptide chain through successive coupling of suitably activatedN-protected amino acids to an amino acid or a C-protected peptide chain,with isolation of the intermediates, subsequent selective deprotectionof the N and C-terminal ends of said fragments and, where necessary, ofthe side chains, until the desired peptide is obtained; ii) solid-phasesynthesis of the peptide chain from the C-terminal to the N-terminal endon an insoluble polymer medium. The peptide is removed from the resin byhydrolysis with anhydrous hydrofluoric acid or trifluoroacetic acid inthe presence of suitable scavengers.

The peptides according to the invention are active on many types ofhuman and animal tumours, preventing their growth and metastatic spread.

The compounds according to the invention are advantageous, especiallycompared with Metastin and the correlated peptides reported in WO06/001499, in terms of anti-tumoral effect and effective doses. Thepeptides according to the invention induce a far more effective responsein reducing tumours (20% reduction by Metastin derivatives, 70%reduction by the products according to the invention), at lowerconcentrations.

For the proposed therapeutic uses, the peptides according to theinvention can be formulated as such, or in the form of salts, inpharmaceutical compositions for oral, parenteral, topical, spray ortransdermal administration, possibly in association with other activeingredients. The unit doses in humans can vary within a wide range,typically from 0.1 μg to 1 g per dose, and preferably between 0.1 mg and100 mg, which can easily be determined by one skilled in the artaccording to the disorder to be treated, its severity, and the weight,sex and age of the patient.

The following examples illustrate the invention in greater detail.

Example 1

Preparation of PICM1, a compound with formula I wherein: L1 is Ace, X₁and X3 are Arg, X₂ is Glu, and X₄ is Phe-NH₂. The % formula of PICM1 istherefore Ace-Arg-Glu-Arg-Phe-NH₂.

An automatic peptide synthesiser starting with 2.5 g of Rink-amide resin(0.2 meq/g) equal to 0.5 mmol of amine groups is used for the synthesis.The Fmoc group is hydrolysed in two successive phases with 30%piperidine in DMF for 3 min. and 7 min. The following compounds are thenreacted, in the order listed: Fmoc-Phe-OH (0.581 g), Fmoc-Arg(Pbf)-OH(0.974 g), Fmoc-Glu(OtBu)-OH (0.638 g), Fmoc-Arg(Pbf)-OH (0.974 g) andacetic acid (0.090 g).

The duration of the acylations is 1 h; the resin is then washed and thereaction tested with the Kaiser ninhydrin assay. If the response isnegative, the Fmoc group is hydrolysed as described above before thenext amino acid is coupled. All the amino acids are coupled bydissolving 1.5 mmol of amino acid in 4 ml of DMF, and added to thedeprotected resin with a mixture of activators consisting of a solutionof 0.780 g of PyBop in 2 ml of DMF, 0.230 g of HOBT in 2 ml of DMF, and250 ml of DIEA. For the detachment of the peptide from the resin and theconcomitant deprotection of the side chains, the dry resin is placed ina reactor to which 20 ml of a solution of TFA, thioanisole,mercaptoethanol and anisole, in the ratio of 9:0.5:0.3:0.2 in weight, isadded. The reaction mixture is kept under agitation for 2 h. Thefiltrate is reduced to a small volume and the peptide is extracted byprecipitation with ether. The precipitate is dissolved with water andfreeze-dried. Finally, the peptide is purified by reverse-phasechromatography and characterised by analytical HPLC, using a Vydac C180.46×25 cm column eluted with a linear gradient in acetonitrilecontaining 0.1% (v/v) of trifluoroacetic acid (phase B) against 0.1%(v/v) aqueous trifluoroacetic acid (Phase A), from 5 to 70% in B in 35min at a flow rate of 1 ml/min, with UV detection at 210 nm. Retentiontime (Rt)=11.8 min.; chromatographic purity >99%. FAB-MS: (MH)⁺=650.

Example 2

Preparation of PICM2, a compound with formula I wherein L1 is pGlu, X₁and X3 are Arg, X₂ is Glu and X₄ is Tyr. The % formula of PICM2 istherefore pGlu-Arg-Glu-Arg-Tyr-OH.

An automatic peptide synthesiser starting with 2.5 g ofFmoc-Tyr(tBu)-Novasyn-TGA resin (0.2 meq/g), equal to 0.5 mmol of aminegroups, is used for the synthesis. The Fmoc group is hydrolysed in twosuccessive phases with 30% piperidine in DMF for 3 min and 7 min. Thefollowing amino acids are then reacted in the order listed:Fmoc-Arg(Pbf)-OH (0.974 g), Fmoc-Glu(OtBu)-OH (0.638 g),Fmoc-Arg(Pbf)-OH (0.974 g) and Fmoc-pGlu-OH (0.638 g).

The duration of the acylations is 1 h; the resin is then washed and thereaction tested with the Kaiser ninhydrin assay. If the response isnegative, the Fmoc group is hydrolysed as described above before thenext amino acid is coupled. All the amino acids are coupled bydissolving 1.5 mmol of amino acid in 4 ml of DMF and added to thedeprotected resin with a mixture of activators consisting of a solutionof 0.780 g of PyBop in 2 ml of DMF, 0.230 g of HOBT in 2 ml of DMF, and250 ml of DIEA. For the detachment of the peptide from the resin and theconcomitant deprotection of the side chains, the dry resin is placed ina reactor to which 20 ml of a solution of TFA, thioanisole,mercaptoethanol and anisole, in the ratio of 9:0.5:0.3:0.2 in weight, isadded. The reaction mixture is kept under agitation for 2 h. Thefiltrate is reduced to a small volume and the peptide is extracted byprecipitation with ether. The precipitate is dissolved with water andfreeze-dried. Finally, the peptide is purified by reverse-phasechromatography and characterised by analytical HPLC, using a Vydac C180.46×25 cm column eluted with a linear gradient in acetonitrilecontaining 0.1% (v/v) trifluoroacetic acid (phase B) against 0.1% (v/v)aqueous trifluoroacetic acid (Phase A), from 5 to 70% in B in 35 min ata flow rate of 1 ml/min, with UV detection at 210 nm. Retention time(Rt)=12.8 min; chromatographic purity >99%. FAB-MS: (MH)⁺=589.

Example 3

Sequences and characterisation data of peptides synthesised with themethods described in examples 1 and 2.

Table 1 shows the sequences and characterisation data of a series ofpeptides synthesised with the methods described in example 1, consistingof PICM3 to PICM39 and PICM54 to PICM67, and in example 2, consisting ofPICM40 to PICM53, suitably adapted to the specific sequences accordingto the common peptide synthesis methodologies.

TABLE 1 Examples of the peptide sequences according to the invention andtheir characterisation Name L1 X1 X2 X3 X4 Rt (min) MH+ PICM3 Glu ArgGlu Arg Phe-NH2 12.6 736 PICM4 Ace Arg Glu Arg Tyr-NH2 13.1 666 PICM5Ace Arg Glu Arg Trp-NH2 14.3 689 PICM6 Ace Arg Glu N(Me)Arg Phe-NH2 12.9664 PICM7 Ace Arg Glu N(Me)Arg Tyr-NH2 13.4 680 PICM8 Ace Arg GluN(Me)Arg Trp-NH2 14.6 703 PICM9 pGlu Arg Glu N(Me)Arg Phe-NH2 12.7 732PICM10 pGlu Arg Glu N(Me)Arg Tyr-NH2 13.2 748 PICM11 pGlu Arg GluN(Me)Arg Trp-NH2 14.4 771 PICM12 pGlu Arg Glu Arg Phe-NH2 12.4 718PICM13 pGlu Arg Glu Arg Tyr-NH2 12.8 734 PICM14 pGlu Arg Glu Arg Trp-NH214.0 757 PICM15 Ace Arg Aib Arg Phe-NH2 15.5 606 PICM16 Ace Arg Aib ArgTyr-NH2 15.2 622 PICM17 Ace Arg Aib Arg Trp-NH2 16.4 645 PICM18 Ace-AibArg Aib Arg Phe-NH2 17.3 694 PICM19 Ace Arg Aib N(Me)Arg Phe-NH2 18.3620 PICM20 Ace Arg Aib N(Me)Arg Tyr-NH2 18.1 636 PICM21 Ace Arg AibN(Me)Arg Trp-NH2 19.3 659 PICM22 pGlu Arg Aib N(Me)Arg Phe-NH2 14.5 688PICM23 pGlu Arg Aib N(Me)Arg Tyr-NH2 14.2 704 PICM24 pGlu Arg AibN(Me)Arg Trp-NH2 16.0 727 PICM25 pGlu Arg Aib Arg Phe-NH2 15.4 674PICM26 pGlu Arg Aib Arg Tyr-NH2 15.2 690 PICM27 pGlu Arg Aib Arg Trp-NH216.6 713 PICM28 Ace Arg Ac5c Arg Phe-NH2 18.5 632 PICM29 Ace Arg Ac5cArg Tyr-NH2 18.2 648 PICM30 Ace Arg Ac5c Arg Trp-NH2 19.4 671 PICM31 AceArg Ac5c N(Me)Arg Phe-NH2 19.3 646 PICM32 Ace Arg Ac5c N(Me)Arg Tyr-NH219.1 662 PICM33 Ace Arg Ac5c N(Me)Arg Trp-NH2 20.7 685 PICM34 pGlu ArgAc5c N(Me)Arg Phe-NH2 17.6 714 PICM35 pGlu Arg Ac5c N(Me)Arg Tyr-NH217.4 730 PICM36 pGlu Arg Ac5c N(Me)Arg Trp-NH2 18.4 753 PICM37 pGlu ArgAc5c Arg Phe-NH2 16.2 700 PICM38 pGlu Arg Ac5c Arg Tyr-NH2 16.0 716PICM39 pGlu Arg Ac5c Arg Trp-NH2 17.1 739 PICM40 Ace Arg Glu Arg Phe-OH11.7 651 PICM41 Ace Arg Glu Arg Tyr-OH 11.5 667 PICM42 Ace Arg Glu ArgTrp-OH 12.6 690 PICM43 Ace Arg Glu Arg(Me) Tyr-OH 12.3 681 PICM44 pGluArg Glu Arg(Me) Phe-OH 12.9 733 PICM45 pGlu Arg Glu Arg Trp-OH 14.5 572PICM46 Ace Arg Aib Arg Phe-OH 15.7 607 PICM47 Ace Arg Aib Arg(Me) Phe-OH16.6 621 PICM48 pGlu Arg Aib Arg(Me) Tyr-OH 14.6 705 PICM49 pGlu Arg AibArg Trp-OH 13.5 714 PICM50 Ace Arg Ac5c Arg Phe-OH 19.1 633 PICM51 AceArg Ac5c Arg(Me) Tyr-OH 18.9 663 PICM52 pGlu Arg AcSc Arg(Me) Trp-OH21.1 754 PICM53 pGlu Arg Ac5c Arg Trp-OH 20.3 740 PICM54 Ace N(Me)ArgAib Arg Phe-NH₂ 16.5 618 PICM55 Ace N(Me)Arg Aib N(Me)Arg Phe-NH₂ 21.2632 PICM56 Ace Arg Aib N(Me)Arg Phe-NH₂ 16.6 618 PICM57 Ace Arg Aib Argα(Me)Phe-NH₂ 14.3 618 PICM58 Ace N(Me)Arg Aib Arg α(Me)Phe-NH₂ 15.2 632PICM59 Ace N(Me)Arg Aib N(Me)Arg α(Me)Phe-NH₂ 18.3 646 PICM60 Ace ArgAib N(Me)Arg α(Me)Phe-NH₂ 19.1 632 PICM61 Ace-Aib N(Me)Arg Aib ArgPhe-NH₂ 20.6 703 PICM62 Ace-Aib N(Me)Arg Aib N(Me)Arg Phe-NH₂ 21.5 717PICM63 Ace-Aib Arg Aib N(Me)Arg Phe-NH₂ 20.5 703 PICM64 Ace-Aib Arg AibArg α(Me)Phe-NH₂ 15.3 703 PICM65 Ace-Aib N(Me)Arg Aib Arg α(Me)Phe-NH₂16.2 717 PICM66 Ace-Aib N(Me)Arg Aib N(Me)Arg α(Me)Phe-NH₂ 17.1 731PICM67 Ace-Aib Arg Aib N(Me)Arg α(Me)Phe-NH₂ 17.8 717

Example 4

Dose-dependent inhibition of cell migration exerted by PICM57.

The effect of PICM57 described in example 3 on the cell migration ofhuman fibrosarcoma HT1080 cells was tested. A Boyden chamber equippedwith polycarbonate filters, having pores with a diameter of 8 μm(Nucleopore), free of polyvinylpyrrolidone and coated with 5 μg/ml ofvitronectin (Promega), was used, as reported in the literature (Carrieroet al., Cancer Res. 59, 5307, 1999). 3×10⁴ cells, in serum-free DMEM(Dulbecco Modified Eagle Medium), were deposited in the uppercompartment of the Boyden chamber. The lower chamber was packed withDMEM containing 10% FBS (Foetal Bovine Serum) as a source ofchemotactics, in the presence of increasing concentrations of PICM57.The cells were incubated for 4 h at 37° C. in humidified air containing5% CO₂. After incubation, the cells adhering to the upper surface of thefilter were mechanically removed, and the migrated cells adhering to thelower surface of the filter were fixed in ethanol, stained withhaematoxylin, and counted at 200× in 10 fields/filter chosen at random.The directional migration in response to FBS in the absence of PICM57was taken as 100% and the effect of PICM57 on cell migration wasevaluated in percentage terms. The data represent the average of threeindependent experiments conducted in duplicate. The antagonistic effectof PICM57 on migration of HT1080 cells towards FBS is dose-dependent(Table 2). It begins at concentrations of 1 aM, reaches 50% of itsmaximum value at concentrations of 1 fM, and reaches the maximum effectat concentrations of 10 fM (66% of inhibition).

The mobility induced by FBS in the presence of Metastin[45-54] wasevaluated under the same experimental conditions (Table 3). The datademonstrate that Metastin[45-54] concentrations 1,000 times greater thanPICM57 (10 nM Metastin[45-54] vs. 10 fM PICM57 are required to obtaincomparable levels of inhibition of directional migration towards FBS.

TABLE 2 Inhibiting effect of PICM57 on the directional migration ofhuman fibrosarcoma HT1080 cells induced by 10% FBS, calculated as apercentage of the migrated cells toward 10% FBS alone (considered 100%)PICM57 concentration 1 10 100 1 10 100 1 10 100 1 10 100 0 aM aM aM fMfM fM pM pM pM nM nM nM Cell Migration 100 69 54 63 43 39 35 37 41 33 3732 34 towards 10% FBS (%)

TABLE 3 Comparison of the inhibiting effects of PICM57, PICM1 andMetastin [45-54] on the directional migration of HT1080 cells induced by10% FBS, expressed as a percentage of the migrated cells compared withthe control (10% FBS). Metastin Concentration [45-54] PICM1 PICM57  0100 100 100  1 aM 100 99 70 10 aM 83 70 54 10 fM 73 56 39 10 pM 53 46 3610 nM 56 35 32

Example 5

Inhibition of cell motility induced by PICM1 independently of thespecies or histogenetic origin of the cell lines.

Cell lines of human or murine origin, derived from various tissues, weresubjected to in vitro directional migration tests employing Boydenchambers as described in example 4, and using 10% FBS as chemotacticsource, in the presence/absence of 100 pM PICM1. Directional migrationin response to FBS in the absence of PICM1 was taken as 100%, and theeffect of PICM1 on cell migration was evaluated as a percentage of saidvalue. The data represent the average of two independent experimentsconducted in duplicate. In all the cell lines tested, an inhibitingeffect of PICM1 on cell migration was observed (Table 4). Although theextent of the inhibition differs between the cell lines used, it wasnever less than 50%, and was also observed in murine melanoma B16 cells,which are unresponsive to Metastin (Ohtaki, T., et al. (2001) Nature411, 613-617).

TABLE 4 Inhibiting effect of PICM1 peptide on the motility of varioustumour cell lines of human or murine origin induced by 10% FBS,expressed as a percentage of the migrated cells compared with thecontrol (10% FBS). 10% FBS 100 Origin Name of cell line Histogenesis pMPICM1 Human MCF-7 Breast cancer 37 Human MDA-MB-231 Breast cancer 47Human A431 Skin cancer 49 Human HeLa Cervical adenocarcinoma 48 HumanU937 Monocytic leukaemia 39 Human M14 Melanoma 59 Human HT1080Fibrosarcoma 34 Human Saos2 Osteosarcoma 43 Murine B16 Melanoma 50

Example 6

Inhibition of motility of cells pre-incubated with PICM57.

Human fibrosarcoma HT1080 cells were incubated with DMEM, or with DMEMcontaining 100 pM PICM57, for 0, 15, 30, 60, or 120 min. The cells were:a) washed with PBS (Phosphate Buffered Saline); b) washed for 5 minutesat 23° C. with 50 mM glycine-HCl buffer pH 3.0 to remove the peptidesfrom the cell surface (Carriero. et al. Clin. Cancer Res. 8, 1299-1308,1997), and then with PBS. The cells were re-suspended in DMEM andsubjected to cell migration tests in Boyden chambers as described inexample 4, using 10% FBS as chemoattractant. The results (Table 5) areexpressed as a percentage of the cells that migrated in the absence ofFBS (basal migration), taken as 100%. The data represent the average oftwo independent experiments conducted in duplicate. Simplepre-incubation of the cells at PICM57 drastically reduces their abilityto migrate towards FBS. 15 minutes' exposure is enough for theinhibition to become comparable with that described in example 4. Acidtreatment after exposure of the cells to 100 pM PICM57 for 15 or 30 minentirely abolishes the inhibiting effects of PICM57 on cell motility.

TABLE 5 Inhibiting effect of peptide PICM57 on the motility of humanfibrosarcoma HT1080 cells pre-incubated with PICM57 and exposed to agradient of 10% FBS. Cell Migration (% of basal migration)Pre-incubation Without (min) Acid Washing With Acid Washing 0 100 100 0393 362 15 98 367 30 103 350 60 112 ND 120 120 ND

Example 7

The inhibition exerted by PICM57 on cell motility is mediated by FPR,the receptor with high affinity for fMLP.

The effect of peptide PICM57 on cell migration was tested in a Boydenchamber as described in example 4, using rat basophilic leukaemia cellsRBL-2H3 lacking the receptor with high affinity for the formylatedpeptide of bacterial origin fMLP (N-formyl-Met-Leu-Phe) (Le, Y., Gong,W., Tiffany, H. L., Tumanov, A., Nedospasov, S., Shen, W., Dunlop, N.M., Gao, J. L., Murphy, P. M., Oppenheim, J. J., Wang, J. M. Amyloid(beta)42 activates a G-protein-coupled chemoattractant receptor,FPR-like-1. J. Neurosci. 21, RC123, 2001) 50 μg/ml fibronectin or 10 nMfMLP was used as chemoattractant. The results are expressed as thepercentage of cells that migrated in the absence of FBS (basalmigration), taken as 100%. The data represent the average of twoindependent experiments conducted in duplicate. In agreement with thefindings reported in the literature, RBL-2H3 cells migrate towardsfibronectin but not towards fMLP. The addition of 100 pM PICM57 to thechemotactic gradient does not reduce the fibronectin-dependent cellmigration (Table 6). Conversely, RBL-2H3 cells stably transfected withthe cDNA of FPR (RBL-2H3/ETFR) acquire the ability to migrate in agradient constituted by 10 nM fMLP. The addition to the chemotacticgradient of 100 pM PICM57 does not reduce the fibronectin-dependent cellmigration, but reduces cell migration towards fMLP to basal levels(Table 6). The definitive demonstration that the target of PICM57 is FPRderives from the observation that in binding assays, the binding of thefluoresceinated peptide formyl-Nle-Leu-Phe-Nle-Tyr-Lys (MolecularProbes) to the surface of the RBL-2H3/ETFR cells is specifically reducedby 60% by pre-incubating the cells with 10 μM PICM57. Pre-incubation ofthe cells with 10 μM Metastin[45-54] does not modify the bond of thefluoresceinated derivative formyl-Nle-Leu-Phe-Nle-Tyr-Lys peptide to thesurface of the RBL-2H3/ETFR cells.

TABLE 6 Inhibition exerted by PICM57 on fMLP-dependent cell motility ofRBL-2H3/ETFR cells. Cell migration (% of basal migration) RBL-2H3RBL-2H3/ RBL-2H3/ETFR Chemotactic RBL-2H3 PICM57 ETFR PICM57 — 100 100100 100 10 nM fMLP 103 102 270 99 50 μg/ml 272 275 220 215 fibronectin

Example 8

Effect of PICM57 on cell proliferation.

Human fibrosarcoma HT1080 cells (1×10³ cells/well) were plated in DMEM10% FBS using 96-well plates in the presence/absence of 100 pM or 100 nMPICM57. At times 24, 48, 72 or 96 h the non-adhering cells were removed,and after repeated rinses with PBS, the cells adhering to the plate werefixed and then stained with a sterile solution containing 1 mg/ml of MTT(3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide, Sigma) for4 h at 37° C. The stain was then removed with 100 μl of dimethylsulphoxide and the corresponding optical density was readspectrophotometrically at 540 nm. The presence of PICM57 at bothconcentrations did not modify the proliferation index (duplication timeapprox. 18 h) of the HT1080 cells at any of the times examined (Table7). The same result was obtained in parallel experiments, when themedium was removed and fresh solutions of 10% FBS/PICM57 were added tothe cells every day for 4 consecutive days.

TABLE 7 Effect of peptide PICM57 on FBS-dependent proliferation of humanfibrosarcoma HT1080 cells, evaluated by spectrophotometric opticaldensity reading (λ: 540 nm). FBS/ FBS/100 pM FBS/ FBS/100 nM 100 pMPICM57 100 nM PICM57 reached Time FBS PICM57 reached every PICM57 every24 hours (h) (OD) (OD) 24 h (OD) (OD) (OD) 48 0.5 0.45 0.51 0.53 0.56 720.94 0.92 1.07 1.05 1 96 2.1 2.02 2.3 2.18 2.2

Example 9

Effect of PICM1 and PICM57 on the invasiveness of human fibrosarcomaHT1080 cells.

The in vitro cell invasion tests were conducted with Boyden chambers,using polyvinylpyrrolidone free filters, having pores with a diameter of8 μm (Nucleopore), coated with 70 μg/filter of matrigel and DMEMcontaining 10% FBS as chemoattractant (Silvestri et al. Int. J. Cancer102, 562-571, 2002). Human fibrosarcoma HT1080 cells (2×10⁵cells/sample) were deposited in the upper compartment of the chamber ina serum-free culture medium. The lower compartment of the chambercontained DMEM with the addition of 10% FBS as chemotactic source, andthe peptides to be assayed, tested at the concentration indicated. Thechambers, thus assembled, were placed at 37° C. in a humidifiedenvironment containing 5% CO₂. After 18 hrs, the cells which had crossedthe matrigel and adhered to the lower surface of the filters were fixed,stained and counted. The data represent the average of two independentexperiments conducted in duplicate. The results, set out in Table 8, areexpressed as the percentage of cells that invade the matrigel in thepresence of FBS, but in the absence of peptide (100%). The inhibitingeffect of PICM1 and PICM57 on the invasiveness of HT1080 cells isdose-dependent.

TABLE 8 Dose-response inhibiting effect of PICM1 and PICM57 on invasionby human fibrosarcoma HT1080 cells induced by 10% FBS. Cell invasiontowards 10% FBS (%) Peptide concentration PICM1 PICM57  0 100 100  1 aM100 69 10 aM 101 59 10 fM 62 41 10 pM 55 37 10 nM 44 31

Example 10 Inhibiting Effect of PICM1 and PICM57 on Neo-Angiogenesis InVitro

The in vitro neo-angiogenesis tests were conducted by exploiting theability of endothelial cells to form, in the presence of pro-angiogenicfactors, cords which extend to form a network of microtubules on a layerof polymerised matrigel. For this type of assay, 5×10⁴ HUVEC cells(Human Umbilical Vein Endothelial Cells) per well were plated in 24-wellplates, in which 300 μl of matrigel was left to polymerise in thepresence of 40 ng/ml of VEGF (Vascular Endothelial Growth Factor), usedas pro-angiogenic agent, and the peptides indicated, at differentconcentrations. Tubule formation was evaluated after 18 hours'incubation at 37° C. in 5% CO₂, by counting the number of tubularstructures in at least 5 fields, chosen at random, with an invertedmicroscope. The effect of the peptides on VEGF-dependent tubule-formingactivity is calculated as a percentage of the number of tubularstructures counted in the presence of VEGF (taken as 100%). The data(Table 9) represent the average of two experiments conducted induplicate. The antagonistic effect of PICM1 and PICM57 onneo-angiogenesis in vitro is dose-dependent. This effect begins atconcentrations of aM and reaches the peak action at a concentration ofpM (20% of the number of tubular structures counted); 50% of the maximumeffect is reached at concentrations of fM. The anti-angiogenic effectexerted by 50 nM of endostatin and the low inhibitory effect exerted bymetastin[45-54] are shown by way of comparison.

TABLE 9 Dose-dependent inhibiting effect of PICM1 and PICM57 on theVEGF-dependent tubule-forming activity of HUVEC endothelial cells.VEGF-dependent tubule-forming activity (%) Metastin Peptideconcentration PICM57 PICM1 Endostatin [45-54]  0 100 100 ND 100   1 aM96 104 ND ND 10 aM 75 77 ND ND 10 fM 53 65 ND 90 10 pM 23 30 ND 63 10 nM19 22 30 65

Example 11 Toxicity of PICM57

Acute toxicity tests of PICM57 on the mouse demonstrate an LD50 of 30mg/Kg, clustered in the interval between 28 mg/Kg (all animals alive)and 32 mg/Kg (all animals dead). Acute toxicity tests of PICM57 on therat demonstrate an LD50 of 65 mg/Kg, clustered in the interval between58 mg/Kg (all animals alive) and 70 mg/Kg (all animals dead).

Example 12 Anti-Metastatic Effect In Vivo

The anti-metastatic power of PICM1 was evaluated on an animal model.2×10⁶ human fibrosarcoma HT1080 cells, which cause pulmonary metastasisin the experimental model (Schweinitz A, et al., J. Biol. Chem.279:33613, 2004), were suspended in saline solution and injected intothe caudal vein of twenty 7-week-old CD1 nude mice. 24 hours after theinoculation of the tumour cells, groups of 10 animals were treated witha slow injection of 3 mg/Kg of PICM1, dissolved in 100 μl of salinesolution, every 48 h, with the animal immobilised. The 10 controlanimals received the same injection treatment, but with the salinesolution only. Each animal was weighed and inspected for dyspnoea everyday. The control animals showed clear signs of respiratory difficultyafter 22 days. All the treated and control animals were sacrificed, andan autopsy was conducted. The lungs, liver, kidneys, spleen and heartwere subjected to macroscopic and microscopic analysis. None of theorgans examined, of the controlled or treated animals, presented anyhisto-morphological alterations, with the exception of the lungs.However, macroscopic analysis of the lungs indicated extensivesub-pleural neoformations in the control animals, often with necroticand haemorrhagic characteristics, which were not observed in the treatedanimals. At microscopic level, serial sections of the pulmonary biopsiesof the control animals showed that 45% of the parenchymal area onaverage was occupied by metastatic nodules, often perivascular, with acentral necrotic area. Conversely, on microscopic observation theanimals treated with 3 mg/Kg of PICM1 did not exhibit the presence ofmetastasis.

1-16. (canceled)
 17. Peptides and their functionally equivalentderivatives, in salified or non-salified form, with the general formulaL₁-X₁—X₂—X₃—X₄, wherein: L₁ is H, or acyl, Glu, Gln, optionallyN-acylated or N-alkylated and/or Cα-alkylated, Pro, hydroxy-Pro, Azt,Pip, pGlu, optionally N-acylated and/or Cα-alkylated, Aib, Ac3c, Ac4c,Ac5c or Ac6c, optionally N-acylated or N-alkylated; X₁ and X₃, which areequal or different, optionally N-alkylated and/or Cα-alkylated, areselected from Arg, Orn and Lys, optionally guanidinylated, andphenylalanine substituted in the meta or para positions with an amine orguanidine group; X₂ is chosen from among Glu, Lys, optionallyN-alkylated and/or Cα-alkylated, Aib, Ac3c, Ac4c, Ac5c and Ac6c,optionally N-alkylated; X₄ is chosen from among Phe, h-Phe, Tyr, Trp,1-NaI, 2-NaI, h-1-NaI, h-2-NaI, Cha, Chg and Phg, optionally amidatedand/or Cα-alkylated.
 18. Peptides as claimed in claim 17, wherein theacylating and alkylating residues contain 1 to 9 carbon atoms, theamidating C-terminal residue contains 0 to 14 carbon atoms, and theCα-alkylating residue contains 1 to 9 carbon atoms.
 19. Peptides asclaimed in claim 17, wherein the acylating and alkylating residuescontain 1 or 2 carbon atoms, and the amidating C-terminal residuecontains 0 to 5 carbon atoms.
 20. Peptides as claimed in claim 17 withthe sequence: Ace-Arg-Glu-Arg-Phe-NH₂; pGlu-Arg-Glu-Arg-Tyr-OH;Glu-Arg-Glu-Arg-Phe-NH₂; Ace-Arg-Glu-Arg-Tyr-NH₂;Ace-Arg-Glu-Arg-Trp-NH₂; Ace-Arg-Glu-N(Me)Arg-Phe-NH₂;Ace-Arg-Glu-N(Me)Arg-Tyr-NH₂; Ace-Arg-Glu-N(Me)Arg-Trp-NH₂;pGlu-Arg-Glu-N(Me)Arg-Phe-NH₂; pGlu-Arg-Glu-N(Me)Arg-Tyr-NH₂;pGlu-Arg-Glu-N(Me)Arg-Trp-NH₂; pGlu-Arg-Glu-Arg-Phe-NH₂;pGlu-Arg-Glu-Arg-Tyr-NH₂; pGlu-Arg-Glu-Arg-Trp-NH₂;Ace-Arg-Aib-Arg-Phe-NH₂; Ace-Arg-Aib-Arg-Tyr-NH₂;Ace-Arg-Aib-Arg-Trp-NH₂; Ace-Aib-Arg-Aib-Arg-Phe-NH₂;Ace-Arg-Aib-N(Me)Arg-Phe-NH₂; Ace-Arg-Aib-N(Me)Arg-Tyr-NH₂;Ace-Arg-Aib-N(Me)Arg-Trp-NH₂; pGlu-Arg-Aib-N(Me)Arg-Phe-NH₂;Glu-Arg-Aib-N(Me)Arg-Tyr-NH₂; pGlu-Arg-Aib-N(Me)Arg-Trp-NH₂;pGlu-Arg-Aib-Arg-Phe-NH₂; pGlu-Arg-Aib-Arg-Tyr-NH₂;pGlu-Arg-Aib-Arg-Trp-NH₂; Ace-Arg-Ac5c-Arg-Phe-NH₂;Ace-Arg-Ac5c-Arg-Tyr-NH₂; Ace-Arg-Ac5c-Arg-Trp-NH₂;Ace-Arg-Ac5c-N(Me)Arg-Phe-NH₂; Ace-Arg-Ac5c-N(Me)Arg-Tyr-NH₂;Ace-Arg-Ac5c-N(Me)Arg-Trp-NH₂; pGlu-Arg-Ac5c-N(Me)Arg-Phe-NH₂;pGlu-Arg-Ac5c-N(Me)Arg-Tyr-NH₂; pGlu-Arg-Ac5c-N(Me)Arg-Trp-NH₂;pGlu-Arg-Ac5c-Arg-Phe-NH₂; pGlu-Arg-Ac5c-Arg-Tyr-NH₂;pGlu-Arg-Ac5c-Arg-Trp-NH₂; Ace-Arg-Glu-Arg-Phe-OH;Ace-Arg-Glu-Arg-Tyr-OH; Ace-Arg-Glu-Arg-Trp-OH;Ace-Arg-Glu-N(Me)Arg-Tyr-OH; pGlu-Arg-Glu-N(Me)Arg-Phe-OH;pGlu-Arg-Glu-Arg-Trp-OH; Ace-Arg-Aib-Arg-Phe-OH;Ace-Arg-Aib-N(Me)Arg-Phe-OH; pGlu-Arg-Aib-N(Me)Arg-Tyr-OH;pGlu-Arg-Aib-Arg-Trp-OH; Ace-Arg-Ac5c-Arg-Phe-OH;Ace-Arg-Ac5c-N(Me)Arg-Tyr-OH; pGlu-Arg-Ac5c-N(Me)Arg-Trp-OH;pGlu-Arg-Ac5c-Arg-Trp-OH; Ace-N(Me)Arg-Aib-Arg-Phe-NH₂;Ace-N(Me)Arg-Aib-N(Me)Arg-Phe-NH₂; Ace-Arg-Aib-N(Me)Arg-Phe-NH₂;Ace-Arg-Aib-Arg-α(Me)Phe-NH₂; Ace-N(Me)Arg-Aib-Arg-α(Me)Phe-NH₂;Ace-N(Me)Arg-Aib-N(Me)Arg-α(Me)Phe-NH₂;Ace-Arg-Aib-N(Me)Arg-α(Me)Phe-NH₂; Ace-Aib-N(Me)Arg-Aib-Arg-Phe-NH₂;Ace-Aib-N(Me)Arg-Aib-N(Me)Arg-Phe-NH₂; Ace-Aib-Arg-Aib-N(Me)Arg-Phe-NH₂;Ace-Aib-Arg-Aib-Arg-α(Me)Phe-NH₂; Ace-Aib-N(Me)Arg-Aib-Arg-α(Me)Phe-NH₂;Ace-Aib-N(Me)Arg-Aib-N(Me)Arg-α(Me)Phe-NH₂;Ace-Aib-Arg-Aib-N(Me)Arg-α(Me)Phe-NH₂.
 21. Pharmaceutical compositionscomprising one of the compounds claimed in claim 17, in association withsuitable vehicles or excipients.
 22. A method for the prevention andtreatment of disorders associated with altered cell migration comprisingadministering an effective amount of a compound of claim 17 to a humanin need thereof.
 23. Method as claimed in claim 22, wherein saideffective amount is from 0.1 μg to 1 g per dose.
 24. Method as claimedin claim 22, wherein said effective amount is between 0.1 mg and 100 mgper dose.
 25. Method as claimed in claim 22 for the prevention andtreatment of local and metastatic invasion by malignant tumours. 26.Method as claimed in claim 22, for the prevention and treatment ofdisorders associated with neo-angiogenesis, including retinalvasculopathies, retinopathies, macular degeneration and oedema, andKaposi's sarcoma.
 27. Method as claimed in claim 22, for the preventionand treatment of disorders supported by cell migration and associatedwith chronic inflammation, including autoimmune diseases, rheumatoidarthritis, psoriasis, and chronic granulomatous disorders.
 28. Method asclaimed in claim 22, for the prevention and treatment of diseasesassociated with herpes virus infection.